P2-195 Effect of Growth Media on Bacterial Pressure Resistance of Escherichia coli K12 lux Bioreporters

Tuesday, August 2, 2016
America's Center - St. Louis
Eileen Duarte Gomez, Purdue University, West Lafayette, IN
Shiyu Cai, Purdue University, West Lafayette, IN
Laszlo Csonka, Purdue University, West Lafayette, IN
Mark Morgan, University of Tennessee-Knoxville, Knoxville, TN
Fernanda San Martin-Gonzalez, Purdue University, West Lafayette, IN
Bruce Applegate, Purdue University, West Lafayette, IN
Introduction: Hydrostatic pressurization has revolutionized traditional food preservation as its application on foodstuffs results in inactivation of undesirable microorganisms. The lack of understanding about the microbial metabolic changes occurring under pressure however has hindered the widespread use of this technology. In addition, pressure induced microbial inactivation seems to be strain and environment dependent.

Purpose: The purpose of this study was to evaluate the effect of nutrient rich and nutrient deficient growth media on pressure resistance of Escherichia coli K12 lux.

Methods: Escherichia coli K12 lux strains were grown in 100 ml nutrient rich Luria-Bertani broth and nutrient deficient M9 media, incubated for 16 hours at 37°C and sub-cultured in 100 ml fresh LB broth and M9 media until an OD600 ~0.5 was reached. A decanal solution was added to 1 ml culture for a 0.001% final concentration. Sample triplicates were contained in 350 μl polypropylene tubes and exposed to 50, 100 and 150 MPa over three 10 minutes compression and 10 minutes decompression cycles during which luciferase expression was recorded using a photomultiplier-tube directly connected to a sapphire window in the pressure vessel.

Results: Although Escherichia coli K12 lux grown in LB media showed slightly higher bioluminescence values than cells grown in M9 media, there was no significant effect of growth media on luciferase expression. Nevertheless, growth conditions had an effect (P < 0.05) on pressure induced bacterial inactivation, as cells grown in M9 media showed 0±0 log reductions for all pressure levels compared to 2±0 and 4±1 log reductions for 100 and 150 MPa for cells grown in LB media.

Significance: These findings suggest that prior exposure to nutrient deficient M9 growth media might have triggered a bacterial stress response that conferred cross protection against pressure. This needs to be considered for an effective and reliable use of this technology for food preservation.